High-efficiency gene electrotransfer into skeletal muscle: Description and physiological applicability of a new pulse generator

Laboratoire de Physiologie, Groupe Physiologie et Physiopathologie de l'Exercice et du Handicap, Groupement d'Intérêt Public-Exercice Sport Santé, Faculté de Médecine J. Lisfranc, 15 rue Ambroise Paré, Saint-Etienne Cedex, France.
Biochemical and Biophysical Research Communications (Impact Factor: 2.3). 09/2002; 296(2):443-50. DOI: 10.1016/S0006-291X(02)00901-4
Source: PubMed


Efficiency and reproducibility of gene electrotransfer depend on the electrical specifications provided by the pulse generator, such as pulse duration, pulse number, pulse frequency, pulse combination, and current intensity. Here, we describe the performances of GET42, a pulse generator specifically designed for gene electrotransfer into skeletal muscle. Expression of beta-galactosidase in the Tibialis anterior muscle of Sprague-Dawley male rats was increased 250-fold by GET42 compared to DNA injection alone. Combination of high and low current intensity pulses further increased transfection efficiency (400-fold compared to DNA injection without electrotransfer). Varying degrees of muscle necrosis were observed after gene electrotransfer. Nevertheless, muscle necrosis was dramatically reduced after optimization of cumulated pulse duration without significant reduction in transfection efficiency. Physiological applicability was illustrated by the analysis of cytochrome c promoter transactivation. In conclusion, GET42 has proven to be a reliable and efficient pulse generator for gene electrotransfer experiments, and provides a powerful mean to study in vivo the regulation of gene expression.

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Available from: Anne-Cécile Durieux, Jun 11, 2014
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    • "Seven days after gene electrotransfer, the animals were anesthetized and the tibialis anterior muscles were removed. Protein isolation and measurement of firefly luciferase activity were performed as described previously [45]. To correct for interindividual variations in transfection efficiency, luciferase activity was normalized to β-galactosidase activity [45]. "
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    • "Indeed, an efficient transfection into skeletal muscles and tumors required the application of limited number of long pulses (several ms) at a low frequency (1 Hz). But, a very high number of repetitive short but stronger pulses (several μs) at a high frequency (kHz) give a high level of gene expression into the same tissues (Lucas et al. 2002; Vicat et al. 2000; Rizzuto et al. 1999; Mir et al. 1999; Durieux et al. 2002). A suitable protocol using a short high voltage pulse (kV/cm, μs) followed by several longer low voltage pulses (V/cm, ms) at a low frequency (1 Hz) has been proposed to aid gene delivery into skeletal muscle (Bureau et al. 2000). "
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